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Development and Clinical Application of a Rapid and Sensitive Loop-Mediated Isothermal Amplification Test for SARS-CoV-2 Infection.
Hu, Xuejiao; Deng, Qianyun; Li, Junmin; Chen, Jierong; Wang, Zixia; Zhang, Xiqin; Fang, Zhixin; Li, Haijian; Zhao, Yunhu; Yu, Pan; Li, Wenmin; Wang, Xiaoming; Li, Shan; Zhang, Lei; Hou, Tieying.
  • Hu X; Division of Laboratory Medicine, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, People's Republic of China.
  • Deng Q; Division of Laboratory Medicine, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, People's Republic of China.
  • Li J; MOE International Joint Lab for Synthetic Biology and Medicine, School of Biology and Biological Engineering, South China University of Technology, Guangzhou, People's Republic of China.
  • Chen J; Division of Laboratory Medicine, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, People's Republic of China.
  • Wang Z; Division of Laboratory Medicine, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, People's Republic of China.
  • Zhang X; Division of Laboratory Medicine, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, People's Republic of China.
  • Fang Z; Department of Laboratory Medicine and Central Laboratories, Guangdong Second Provincial General Hospital, Guangzhou, People's Republic of China.
  • Li H; Department of Laboratory Medicine and Central Laboratories, Guangdong Second Provincial General Hospital, Guangzhou, People's Republic of China.
  • Zhao Y; Division of Laboratory Medicine, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, People's Republic of China.
  • Yu P; Genskey Biotechnology Co. Ltd., Beijing, People's Republic of China.
  • Li W; Division of Laboratory Medicine, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, People's Republic of China.
  • Wang X; MOE International Joint Lab for Synthetic Biology and Medicine, School of Biology and Biological Engineering, South China University of Technology, Guangzhou, People's Republic of China.
  • Li S; MOE International Joint Lab for Synthetic Biology and Medicine, School of Biology and Biological Engineering, South China University of Technology, Guangzhou, People's Republic of China.
  • Zhang L; MOE International Joint Lab for Synthetic Biology and Medicine, School of Biology and Biological Engineering, South China University of Technology, Guangzhou, People's Republic of China lzhangce@scut.edu.cn houtieying@gdph.org.cn.
  • Hou T; Division of Laboratory Medicine, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, People's Republic of China lzhangce@scut.edu.cn houtieying@gdph.org.cn.
mSphere ; 5(4)2020 08 26.
Artículo en Inglés | MEDLINE | ID: covidwho-730989
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ABSTRACT
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) outbreak urgently necessitates sensitive and convenient COVID-19 diagnostics for the containment and timely treatment of patients. We aimed to develop and validate a novel reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay to detect SARS-CoV-2. Patients with suspected COVID-19 and close contacts were recruited from two hospitals between 26 January and 8 April 2020. Respiratory samples were collected and tested using RT-LAMP, and the results were compared with those obtained by reverse transcription-quantitative PCR (RT-qPCR). Samples yielding inconsistent results between these two methods were subjected to next-generation sequencing for confirmation. RT-LAMP was also applied to an asymptomatic COVID-19 carrier and patients with other respiratory viral infections. Samples were collected from a cohort of 129 cases (329 nasopharyngeal swabs) and an independent cohort of 76 patients (152 nasopharyngeal swabs and sputum samples). The RT-LAMP assay was validated to be accurate (overall sensitivity and specificity of 88.89% and 99.00%, respectively) and diagnostically useful (positive and negative likelihood ratios of 88.89 and 0.11, respectively). RT-LAMP showed increased sensitivity (88.89% versus 81.48%) and high consistency (kappa, 0.92) compared to those of RT-qPCR for SARS-CoV-2 screening while requiring only constant-temperature heating and visual inspection. The time required for RT-LAMP was less than 1 h from sample preparation to the result. In addition, RT-LAMP was feasible for use with asymptomatic patients and did not cross-react with other respiratory pathogens. The developed RT-LAMP assay offers rapid, sensitive, and straightforward detection of SARS-CoV-2 infection and may aid the expansion of COVID-19 testing in the public domain and hospitals.IMPORTANCE We developed a visual and rapid reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay targeting the S gene for SARS-CoV-2 infection. The strength of our study was that we validated the RT-LAMP assay using 481 clinical respiratory samples from two prospective cohorts of suspected COVID-19 patients and on the serial samples from an asymptomatic carrier. The developed RT-LAMP approach showed an increased sensitivity (88.89%) and high consistency (kappa, 0.92) compared with those of reverse transcription-quantitative PCR (RT-qPCR) for SARS-CoV-2 screening while requiring only constant-temperature heating and visual inspection, facilitating SARS-CoV-2 screening in well-equipped labs as well as in the field. The time required for RT-LAMP was less than 1 h from sample preparation to the result (more than 2 h for RT-qPCR). This study showed that the RT-LAMP assay was a simple, rapid, and sensitive approach for SARS-CoV-2 infection and can facilitate COVID-19 diagnosis, especially in resource-poor settings.
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Texto completo: Disponible Colección: Bases de datos internacionales Base de datos: MEDLINE Asunto principal: Neumonía Viral / Infecciones por Coronavirus / Técnicas de Laboratorio Clínico / Técnicas de Amplificación de Ácido Nucleico / Técnicas de Diagnóstico Molecular / Betacoronavirus Tipo de estudio: Estudio de cohorte / Estudios diagnósticos / Estudio observacional / Estudio pronóstico / Ensayo controlado aleatorizado Límite: Adulto / Femenino / Humanos / Masculino / Middle aged Idioma: Inglés Año: 2020 Tipo del documento: Artículo

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Texto completo: Disponible Colección: Bases de datos internacionales Base de datos: MEDLINE Asunto principal: Neumonía Viral / Infecciones por Coronavirus / Técnicas de Laboratorio Clínico / Técnicas de Amplificación de Ácido Nucleico / Técnicas de Diagnóstico Molecular / Betacoronavirus Tipo de estudio: Estudio de cohorte / Estudios diagnósticos / Estudio observacional / Estudio pronóstico / Ensayo controlado aleatorizado Límite: Adulto / Femenino / Humanos / Masculino / Middle aged Idioma: Inglés Año: 2020 Tipo del documento: Artículo